Preparation method of multidimensional multivalence prostatic cancer specificity tumor antigen assembly

A technology for prostate cancer and tumor antigen, applied in the field of biomedicine preparation, can solve the problems of low immune killing and poor treatment effect, and achieve the effect of long stimulation time, inhibiting and killing prostate cancer, and good effect.

Active Publication Date: 2019-12-03
天津市泌尿外科研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to solve the problem of low immune killing and poor therapeutic effect caused by the use of prostate specific antigen pe...

Method used

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  • Preparation method of multidimensional multivalence prostatic cancer specificity tumor antigen assembly
  • Preparation method of multidimensional multivalence prostatic cancer specificity tumor antigen assembly
  • Preparation method of multidimensional multivalence prostatic cancer specificity tumor antigen assembly

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Materials and methods

[0025] 1.1.1 Strains: The strains used are derived from Mycobacterium bovis (BCG, BCG; Pasteur strain) provided by ATCC (authorized by ATCC and signed a development and use agreement).

[0026] 1.1.2 Culture medium: purchased from sigma company.

[0027] 1.1.3 Nano-polymer materials:

[0028] Chitosan, polyglutamic acid, etc.; purchased from Tianjin Zhongao Tianyuan Technology Development Co., Ltd.

[0029] 1.1.4 General chemical reagents: (1) EDS (ethylenediaminetetraacetic acid), NHSS (N-hydroxysulfosuccinic acid amine); (2) sulfate; (3) organic acid; (4) inorganic salt, etc.;

[0030] All were purchased from Tianjin Sino-Australian Tianyuan Technology Development Co., Ltd.

[0031] 1.1.5 Prostate cancer tumor markers PSMA, PAP, STEAP1, and ACPP protein coding sequences were synthesized by Tianjin "Jinweizhi" company with reference to the standards published by "NCBI".

[0032] 1.1.6 Cloning Pichia pastoris strains and plasmids were used...

Embodiment 2

[0044]1. Modification and synthesis of graphene oxide (GO) modified polypeptide:

[0045] Take 10 mL of an aqueous solution made of nanographene oxide (GO), (1 mg / mL), add EDS (5 mg) and NHSS (4 mg) into the GO aqueous solution, and stir for 30 min. Then 3 mL (1 mg / mL) of each (PSMA, PSCA, PAP or STEAP) polypeptide aqueous solution was added, and the reaction was stirred at room temperature for 48 h. The reaction solution was centrifuged, the supernatant was discarded, the obtained precipitate was washed with water three times, and the precipitate was collected by centrifugation. It was stirred in a weakly acidic aqueous solution for 72 hours at a low speed, and re-dispersed in 10 mL of deionized water to obtain a graphene oxide (GO) modified polypeptide stock solution. and stored at 4°C. 2. Graphene oxide (GO) - protein connection modification:

[0046] Take 2 mL of graphene oxide (GO) modified polypeptide stock solution, add it to the separated and purified BCG bacterial ...

Embodiment 3

[0048] 1. Polyglutamic acid-polypeptide modification solution.

[0049] Dilute each (PSMA, PSCA, PAP or STEAP) polypeptide stock solution (1mg / mL) with PBS solution, add to polyglutamic acid containing 5% acetic acid dispersion solution (1mg / mL), the ratio is set at 3:1~ 6:1, vortex, mix thoroughly, place at 4°C, stir at low speed, overnight. The prepared polyglutamic acid-polypeptide solution was obtained.

[0050] 2. Polyglutamic acid polypeptide-protein connection modification:

[0051] Take 2 mL of the prepared polyglutamic acid-polypeptide liquid peptide stock solution, add it to the separated and purified BCG bacterial protein PBS solution (containing 2 mg / 8 mL), and adjust the pH to neutral. Add EDS and NHSS and stir for 30 minutes to fully dissolve. Adjust the pH to 7.0, stir at a low speed at 20°C, and react with stirring for 24 hours. After the reaction, the "assembly" solution was obtained, which was stored at 4°C for future use.

[0052] 1.4. Electrophoretic m...

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Abstract

The invention relates to a preparation method of a multidimensional multivalence prostatic cancer specificity tumor antigen assembly. Cultured mycobacterium bovis(BCG vaccine BCG) thalli, and throughseparation, refining and purifying, obtained thalli protein is a basic carrier. The effect of the multidimensional multivalence prostatic cancer specificity tumor antigen assembly can mobilize and activate an immune response reaction of an organism autoimmunity system. A nanometer polymer is used as a multi-connection arm, and self modified structure functional groups are utilized. Through a combination manner of covalent bonds or non-covalent bonds, thalli protein and specific prostatic cancer and tumor antigen polypeptide genetic fragments are connected. The multidimensional multivalence three-dimensional configuration bridge linkage mode is finished, and the multidimensional multivalence prostatic cancer specificity tumor antigen assembly having the characteristics of being high in biological safety, clear in specific antigen, high in autosensitization original nature and the like is prepared. The preparation method can be used for preparing a prostatic cancer specificity antibody,is used for pinpoint research, development and production of prostatic cancer vaccines for human, and has favorable wide application prospects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine preparation. Background technique [0002] Prostate cancer is one of the main malignant cancers that threaten the life safety of elderly men. In the United States, prostate cancer is the second largest tumor killer for men. With the improvement of living standards in my country, the incidence of prostate cancer in China has gradually increased, and it currently ranks 10th in the world. The treatment methods for prostate cancer include surgical treatment, androgen deprivation therapy (castration therapy), active monitoring, etc. However, after a period of castration therapy, it turns into hormone-independent prostate cancer, resulting in the death of the patient. Therefore, at present we need to find highly effective prostate cancer control and treatment strategies. [0003] In recent years, immunotherapy is considered to be one of the most likely ways to conquer tumors. With the advancement...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K47/64A61P35/00
CPCA61K39/001195A61K47/64A61P35/00A61K2039/70A61K2039/6068A61K2039/627A61K2039/64A61K2039/884
Inventor 牛远杰陈家童林平生喻其林杨洋国林沛蔡启亮尚芝群唐慧琴
Owner 天津市泌尿外科研究所
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